Changes in Intensity and Spectral Distribution of Fluorescence: Effect of Light Treatment on Normal and DCMU*-Poisoned Anacystis nidulans‡

The intensity of the "steady-state" fluorescence of "aerobic" Anacystis nidulans is variable under prolonged illumination with orange (590 mmu) or blue (440 mmu) light for both normally photosynthesizing and DCMU-poisoned cells. In general, orange light illumination causes an increase of the fluorescence intensity followed by a decrease, while blue light causes an increase until a steady level is reached. Poisoned Anacystis cells show four to eight times larger changes in fluorescence intensity than the normal cells; the detailed time course of fluorescence changes is also different in poisoned and normal cells. When algae are cooled to -196 degrees C in light, the light-induced changes in the "steady-state" fluorescence disappear in both types of cells. Difference fluorescence spectra, constructed by subtracting the fluorescence spectra taken after 5-15 min of illumination from those after 60-90 min of illumination, show a doublet structure of the difference band with a major peak coinciding with the Anacystis emission maximum (685 mmu) and a minor peak located at about 693 mmu.     

Control of excitation transfer in photosynthesis. IV. Kinetics of chlorophyll a fluorescence in Porphyra yezoensis

The kinetics of chlorophyll a fluorescence were measured at 685 nm in intact cells of Porphyra yezoensis during alternate illumination of the organism with two colors of light, one absorbed by phycoerythrin and the other by chlorophyll a. Two components of fluorescence change overlapping each other in time were separated; the fast component may be controlled by the rate of Photoreaction II which competes with the fluorescence emission process, and the slow component by the light-induced change in excitation transfer between two pigment systems as suggested in our previous study⁶. The kinetics of the slow change in fluorescence yield were extensively investigated.

Terms, “State I” and “State II” are used to describe the state of excitation transfer. In the State I a lesser amount of excitation energy is delivered in Pigment System I and greater to Pigment System II than in the State II. The conversion of the states is achieved by the selective illumination of pigment systems.

The conversion from the State I toward the State II occurred under Light II (light absorbed by Pigment System II) with a half time of about 10 sec, and it saturated at a light intensity of less than 1000 ergs×cm⁻²×sec⁻¹. The reverse conversion occurred under Light I (light absorbed by Pigment System I) with a half time of about 5 sec, and it saturated at about 10 000 ergs×cm⁻²×sec⁻¹.

Light I and Light II competed with each other in the interconversion of the states.     

Transfer of the Excitation Energy in Anacystis nidulans Grown to Obtain Different Pigment Ratios

The blue-green alga, Anacystis nidulans, was grown in lights of different colors and intensities, and its absorption and fluorescence properties were studied. Strong orange light, absorbed mainly by phycocyanin, causes reduction in the ratio of phycocyanin to chlorophyll a; strong red light, absorbed mainly by chlorophyll, causes an increase in this ratio. This confirms the earlier findings of Brody and Emerson (12) on Porphyridum, and of Jones and Myers (8) on Anacystis. Anacystis cultures grown in light of low intensity show, upon excitation of phycocyanin, emission peaks at 600 mmu and 680 mmu, due to the fluorescence of phycocyanin and chlorophyll a, respectively. Changes in the efficiency of energy transfer from phycocyanin to chlorophyll a are revealed by changes in the ratios of these two bands. A decrease in efficiency of energy transfer from phycocyanin to chlorophyll a seems to occur whenever the ratio of chlorophyll a to phycocyanin deviates from the normal. Algae grown in light of high intensity show, upon excitation of phycocyanin, only a fluorescence band at 660 mmu and no band at 680 mmu. This suggests reduced efficiency of energy transfer from phycocyanin to the strongly fluorescent form of chlorophyll a (chlorophyll a(2)) and perhaps increased transfer to the weakly fluorescent form of chlorophyll a (chlorophyll a(1)).     

Quenching of a room temperature fluorescence band at 693 nm during photoactivation of the water-splitting system of Photosystem II in flashed barley leaves

The modifications of the room temperature fluorescence spectrum during the photoactivation of the water-splitting system by continuous illumination were investigated in flashed barley leaves. A blue shift of the chlorophyll fluorescence band was detected during the first 2 min of illumination. During this shift, a decrease of the fluorescence intensity around 693 nm could be demonstrated in difference spectra and in second derivative spectra. This decrease is interpreted as a quenching of PS II fluorescence during the photoactivation. A relative fluorescence increase around 672 nm also occurred during the same period and is thought to reflect rapid light-induced chlorophyll formation. The flashed leaves contained small amounts of photoactive photochlorophyllide which could be removed by a short flash of intense white light given before continuous illumination. The fact that such flash had only weak effect on the 693 nm fluorescence decrease, whereas it strongly reduced the amplitude of the 672 nm fluorescence increase, favours the above interpretations.Abbreviations chl chlorophyll - PS II Photosystem II - PS I Photosystem I     

Thylakoid protein phosphorylation during State 1—State 2 transitions in osmotically shocked pea chloroplasts

In osmotically shocked pea chloroplasts illuminated with modulated blue-green light (light 2), phosphorylation of the light-harvesting chlorophyll $\text{a}\text{b-}\text{protein}$ complex (LHCP) accompanies the slow decrease in modulated fluorescence that indicates adaptation to light absorbed predominantly by Photosystem II (State 2). On subsequent additional illumination with continuous far-red light (absorbed predominantly by Photosystem I; light 1) both effects are reversed: modulated chlorophyll fluorescence emission increases (indicating adaptation towards State 1) and LHCP is dephosphorylated. Net phosphorylation and dephosphorylation of LHCP induced by light 2 and excess light 1, respectively, occur on the same time scale as the ATP-dependent chlorophyll fluorescence changes indicative of State 2 and State 1 transitions. The phosphatase inhibitor NaF (10 mM), stimulates the effect of blue-green light on fluorescence and prevents the effect of far-red light. These results provide a demonstration that light of different wavelengths can control excitation energy distribution between the two photosystems via the plastoquinol-activated LHCP phosphorylation mechanism suggested previously (Allen, J.F., Bennett, J., Steinback, K.E. and Arntzen, C.J. (1981) Nature 291, 25–29; and Horton, P. and Black, M.T. (1980) FEBS Lett. 119, 141–144).     

Excitation-energy distribution in green algae. The existence of two independent light-driven control mechanisms

Three distinct states can be identified for cells of the green alga Chlorella vulgaris; State 1 and State 2 obtained by preillumination in far-red and red light, respectively, and the dark state obtained by dark-adaptation. Addition of the inhibitor DCMU to algal cells leads to an initial rapid increase in chlorophyll-a fluorescence reflecting the closure of Photosystem II traps. This, in the case of dark and state-2-adapted algae is followed by a slow light-dependent increase to a fluorescence yield typical of State-1-adapted cells. Measurements of low temperature (77 K) emission spectra indicate that the low fluorescence yields of dark and State-2-adapted algae reflect similar balances in excitation-energy distribution between the two photosystems. In both cases, the balance favours PS I and the slow fluorescence increase seen in the poisoned algae reflects a redressing of this balance in favour of PS II. The low fluorescence yield of State-2-adapted algae is thought to be associated with the phosphorylation of chlorophyll a/b light-harvesting protein (Biochim. Biophys. Acta (1983) 724, 94–103). Measurements of the uncoupler and ATPase sensitivity of the light-dependent increases seen in DCMU-poisoned cells indicate that the low fluorescence yield of dark-adapted algae is of different origin. Evidence is presented showing that the light-driven changes in excitation-energy distribution seen in green algae involve two distinct processes; a low-intensity, wavelenght-independent change reflecting simple light/dark changes and a higher intensity, wavelength-dependent change reflecting State 1/State 2 adaptation. The former changes appear to be associated with changes in the local ionic environment within the algal chloroplast, whilst the latter appear to reflect changes in the phosphorylation state of chlorophyll a/b light-harvesting protein.     

Regulation of cyanobacterial photosynthesis determined from variable fluorescence yields of photosystem II

Abstract The cyanobacteria Fremyella diplosiphon 7601 and Synechocystis 6701 were grown in continuous cultures with monochromatic red light (680 nm). The distribution of light energy over photosystem I and II was determined from changes in PS II fluorescence at 685 nm. In both organisms, wavelengths absorbed primarily by chlorophyll a caused the high fluorescent state of PS II (State 1), while wavelengths absorbed by the phycobilisome led to low PS II fluorescence (State 2). Superimposing continuous light 2 on the excitation light yielded State 2 fluorescence patterns for Synechocystis 6701, while F. diplosiphon 7601 showed fluorescence patterns similar to state 1 → 2 transitions and changes in fluorescence yield were related to the intensity of the background light. Some ecological implications of energy (re)distribution in cyanobacterial photosynthesis are discussed.     

Energization and ultrastructural pattern of thylakoids formed under periodic illumination followed by continuous light

Bean leaves grown under periodic illumination (56 cycles of 2 min light and 98 min darkness) were subsequently exposed to continuous illumination, and in connection with granum formation and accumulation of the light-harvesting pigment-protein complex thermoluminescence and light-induced shrinkage of thylakoid membranes were studied. Juvenile chloroplasts with large double sheets of thylakoids obtained under periodic light exhibited low temperature spectra of polarized fluorescence yielding fluorescence polarization (FP) values < 1 at 695 nm, characteristic for pheophytin emission. In the course of maturation under continuous light when normal grana appeared and the chlorophyll a/b light-harvesting photosystem II complex was incorporated into the membrane, at 695 nm the relative intensity of fluorescence dropped and FP changed to a value of > 1, suggesting an overlap between the emission of pheophytin and that of the chlorophyll a/b light-harvesting photosystem II complex. Thermoluminescence glow curves recorded with juvenile thylakoids displayed a relatively high proportion of emission at low temperatures (around -10°C) while with mature chloroplasts, more thermoluminescence originated from energetically deeper traps (discharged around 28°C). This means that during thylakoid development the capacity of the membrane to stabilize the separated charges increases, which might be favourable for the ultimate conservation of energy. The more extensive energization of mature thylakoids was also indicated by a light-induced decrease in the thickness of the membranes upon illumination; a change which could not be detected in juvenile thylakoids.Abbreviations EDTA ethylenediamine tetraacetic acid - Hepes 4-(2-hydroxy ethyl)-1-piperazine ethane sulfonic acid Dedicated to Prof. L.N.M. Duysens on the occasion of his retirement.     

Changes in the distribution of light energy between the two photosystems in spinach leaves

Time courses of chlorophyll fluorescence at room temperature and fluorescence spectra at 77 K were measured to investigate the light-induced changes in the distribution of light energy between the two photosy stems in young spinach leaves. Illumination of the dark adapted leaves with primarily system II light induced typical fluorescence transients at room temperature. Fluorescence spectra at 77 K showed that the intensity of system II fluorescence at 77 K changed nearly in parallel with the fluorescence transients at room temperature within the range from M₁ to T during illumination of the leaf. Illumination of the dark adapted leaves with light I produced an increase of system II fluorescence measured at 77 K. The characteristics of the changes induced by light I or II were different, showing that these two effects are related to different mechanisms. These results suggest that the dark state in spinach leaves is state II, that light I induces a state II to I transition, while light II induces fluorescence changes that are produced by mechanisms other than state I-state II transitions.     

Light-induced changes in the fluorescence yield of chlorophyll a in Anacystis nidulans II. The fast changes and the effect of photosynthetic inhibitors on both the fast and slow fluorescence induction

1. The intensity dependence and spectral variations during thefast transient of chlorophyll a (Chl a) fluorescence have beenanalyzed in the blue-green alga Anacystis nidulans. (Unlikethe case of eukaryotic unicellular green or red algae, the fastfluorescence induction characteristics of the prokaryotic blue-greenalgae had not been documented before.)
2. Dark adapted cellsof Anacystis exhibit two types of fluctuationsin the fluorescenceyield when excited with bright orange light(absorbed mainlyin phycocyanin). The first kinetic patterncalled the fast (sec)fluorescence transient exhibits a characteristicoriginal levelO, intermediary hump I, a pronounced dip D, peakP and a transitorysmall decline to a quasi steady state S.After attaining S,fluorescence yield slowly rises to a maximumlevel M. From M,the decline in fluorescence yield to a terminalT level is extremelyslow as shown earlier by Papageorgiou andGovindjee (8). Ascompared with green and red algae, blue-greenalgae seem tohave a small PS decline and a very characteristicslow SM rise,with a M level much higher than the peak P.
3. A prolonged darkadaptation and relatively high intensity ofexciting illuminationare required to evoke DPS type yield fluctuationsin Anacystis.At low to moderate intensities of exciting light,the time forthe development of P depends on light doses, butfor M, thisremains constant at these intensities.
4. Fluorescence emissionwas heterogeneous during the inductionperiod in Anacystis;the P and the M levels were relativelyenriched in short-wavelengthsystem II Chi a emission as comparedto D and S levels.
5. Thefast DPS transient was found to be affected by electrontransportcofactor (methyl viologen), and inhibitors (e.g.,DCMU, NH₂OH)in a manner suggesting that these changes are mostlyrelatedto the oxido-reduction level of intermediates betweenthe twophotosystems. On the other hand, the slow SM changesin fluorescenceyield, as reported earlier (5, 15), paralleloxygen evolution.These changes were found to be resistant toa variety of electrontransport inhibitors (O-phenanthroline,HOQNO, salicylaldoxime,DCMU, NH₂OH and Antimycin a). It issuggested that, in Anacystis,even in the presence of so-calledinhibitors of cyclic electronflow, a "high energy state" isstill produced.
6. Measurementsof Chlorophyll a fluorescence and delayed lightemission inthe presence of both DCMU and NH₂OH indicate thatthe slow SMchanges are not due to the recovery of the reactioncenter IIin darkness preceeding illumination.
7. Our results, thus, suggestthat in Anacystis a net electrontransport supported oxidation-reductionstate of the quencherQ regulates only partially the developmentof the DPS transient,but the development of the slow fluorescenceyield changes seemsnot to be regulated by these reactions.It appears, from datapresented elsewhere, that the slow risein the yield resultsdue to a structural modification of thethylakoid membrane.

¹We are grateful to the National Science Foundation for financialsupport. (Received November 21, 1972; )     

Evidence against proton gradient formation being the cause of chlorophyll fluorescence quenching by N-methylphenazonium methosulfate

In strong illumination, 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (DCMU)-poisoned chloroplasts exhibit a high yield of chlorophyll fluorescence while $\text{P-700}$ turnover, proton uptake, and phosphorylation are inhibited and a pH gradient is undetectable. When 10 μM $\text{N-}\text{methylphenazonium}$ methosulfate (PMS) is included, the fluorescence yield in light is substantially reduced, and when 100 μM ascorbate is also included, the yield is diminished approximately to the level in darkness. Only very slight increases in $\text{P-700}$ turnover and proton uptake (but no detectable pH gradient) accompany the fluorescence yield decline.When 10 μM PMS and 15 mM ascorbate are added to poisoned chloroplasts (the oxygen concentration being greatly reduced), $\text{P-700}$ turnover, proton uptake, the pH gradient and phosphorylation all reach high levels. In this case, the yield of chlorophyll fluorescence is low and is the same in both light and dark. Further addition of an uncoupler eliminates proton uptake, the pH gradient and phosphorylation but does not significantly elevate the fluorescence yield. From these observations we suggest that, in DCMU-poisoned chloroplasts, the fluorescence quenching with PMS occurs by a mechanism unrelated to the generation of a phosphorylation potential.With chloroplasts unpoisoned by DCMU, PMS quenches fluorescence and considerably stimulates proton uptake, the pH gradient and phosphorylation. However, in this case, PMS serves to restore net electron transport.     

Evidence against proton gradient formation being the cause of chlorophyll fluorescence quenching by N-methylphenazonium methosulfate.

In strong illumination, 3-(3, 4-dichlorophenyl)-1,1-dimethylurea (DCMU)-poisoned chloroplasts exhibit a high yield of chlorophyll fluorescence while P-700 turnover, proton uptake, and phosphorylation are inhibited and a pH gradient is undectectable. When 10muM N-methylphenazonium methosulfate (PMS) is included, the fluorescence yield in light is substantially reduced, and when 100 muM ascorbate is also included, the yield is diminished approximately to the level in darkness. Only very slight increases in P-700 turnover and proton uptake (but no detectable pH gradient) accompany the fluorescence yield decline. When 10muM PMS and 15 mM ascorbate are added to poisoned chloroplasts (the oxygen concentration being greatly reduced), P-700 turnover, proton uptake, the pH gradient and phosphorylation all reach high levels. In this case, the yield of chlorophyll fluorescence is low and is the same in both light and dark. Further addition of an uncoupler eliminates proton uptake, the pH gradient and phosphorylation but does not significantly elevate the fluorescence yield. From these observations we suggest that, in DCMU-poisoned chloroplasts, the fluorescence quenching with PMS occurrs by a mechanism unrelated to the generation of a phosphyorylation potential. With chloroplasts unpoisoned by DCMU, PMS quenches fluorescence and considerably stimulates proton uptake, the pH gradient and phosphorylation. However, in this case, PMS serves to restore net electron transport.     

Hydrogen Photoevolution Indicates an Increase in the Antenna Size of Photosystem I in Chlamydobotrys stellata during Transition from Autotrophic to Photoheterotrophic Nutrition

The changes in the light-harvesting antenna size of photosystem I were investigated in the green alga Chlamydobotrys stellata during transition from autotrophic to photoheterotrophic nutrition by measuring the light-saturation behavior of hydrogen evolution following single turnover flashes. It was found that during autotrophic-to-photoheterotrophic transition the antenna size of photosystem I increased from 180 to 250 chlorophyll. The chlorophyll (a + b)/P700 ratio decreased from 800 to 550. The electron transport of photosystem I measured from reduced 2,6-dichloro-phenolindophenol to methylviologen was accelerated 1.4 times. In the 77K fluorescence spectra, the photosystem II fluorescence yield was considerably lowered relative to the photosystem I fluorescence yield. It is suggested that the increased light-harvesting capacity and redistribution of absorbed excitation energy in favor of photosystem I is a response of photoheterotrophic algae to meet the ATP demand for acetate metabolism by efficient photosystem I cyclic electron transport when the noncyclic photophosphorylation is inhibited by CO₂ deficiency.     

Fluorescence and oxygen evolution from Chlorella pyrenoidosa

The process of photosynthetic energy conversion in Chlorella pyrenoidosa was investigated by simultaneous measurement of transient and steady-state rates of O₂ evolution and fluorescence.

1. 1. Alternation or superimposition of light 1 and light 2 illumination induces both fast and slow changes in fluorescence and rate of O₂ evolution. The fast changes are ascribed to changes in conditions of the reaction centers in the context of the ¹ model and the kinetic analysis of ². The slow changes are interpreted as adaptations to the intensity and wavelength of illumination. The adaptive mechanism is described in terms of slow variation in fraction () of total absorbed quanta delivered to System 2. At low intensities, the calculated value of for cells adapted to light 2 illumination (light 2 state) is approx. 0.9 of for cells adapted to light 1 illumination (light 1 state).

2. 2. An increase in fluorescence yield was found to accompany the decrease in O₂ yield at the onset of light saturation with either light 1 or light 2 excitation. Variation in is proposed to account for the differences between the maximum fluorescence yield observed in steady-state conditions and the 1.5 times higher maximum yield observed in transient conditions or in cells inhibited by 3(3,4-dichlorophenyl)-1,1-dimethylurea. Variation in can also explain the observation of a higher rate of fluorescence emission with light 1 excitation than with light 2 excitation for a given steady-state rate of O₂ evolution.

3. 3. A model for energy conversion by System 2 is proposed to account for our observations. The model proposes competitive dissipation of absorbed energy by photochemical trapping at reaction centers and by fluorescence and radiationless de-excitation from both the pigment bed and reaction centers of System 2.

pH control of the chlorophyll a fluorescence in algae

The pH of the suspension medium was found to have a remarkable influence on the “slow” (min) time course of Chlorophyll a fluorescence yield in the green alga Chlorella pyrenoidosa and in the blue-green alga Anacystis nidulans. In Chlorella, the decay of fluorescence yield, in the 1- to 5-min region, is strongly retarded at alkaline pH; this decay rate shows an optimum at pH 6–7. In Anacystis, the rise of fluorescence yield, in the same time range, is decreased optimally at pH 6–7; poisoning with 3(3,4-dichlorophenyl)-1,1-dimethylurea reverses the direction of this pH effect. These observations suggest a correlation of the H⁺ status (or the processes associated with it such as photophosphorylation and resulting conformational changes) of the chloroplast to the yield of chlorophyll a fluorescence in vivo.     

Photosynthetic Apparatus Formation during the Cell Cycle of Chlorella

Synchronous cell division in cultures of Chlorella vulgaris Beijerinck was induced by intermittent illumination: 9 hours light, 6 hours darkness. The rate of photosynthetic O₂ evolution per cell increases 4-fold in a one-step manner at the beginning of the light period, to the same extent as the increase in cell number. Over the division cycle, the following accumulation times during the light period were found: chlorophyll a, between 2 and 8 hours, chlorophyll b, between 5 and 8 hours, reaction centers of photosystems I and II, between 2 and 6 hours; and cytochrome f, between 2.5 and 5 hours. Cytochrome f accumulation is closely followed by an increase in amplitude of the rapid phase in light-induced absorption increase at 520 nanometers and in intensity of the delayed light emission. Enhancement of the delayed fluorescence yield per flash under continuous illumination (caused by the establishment of the pH difference across the thylakoid membrane) is maximal by the first hour of the light period.     

Light- and temperature-induced changes in the distribution of excitation energy between Photosystem I and Photosystem II in spinach leaves

The influence of light quality and temperature on the distribution of the absorbed quanta between Photosystem I (PS I) and Photosystem II (PS II) in spinach leaves has been studied from the characteristics of chlorophyll fluorescence at 77 K. Leaves were preilluminated at different temperatures with either PS I light (to establish State 1) or with PS II light (to establish State 2), then cooled to 77 K and measured for fluorescence. In State 1, energy distribution appeared to be unaffected by temperature. A transition to State 2 resulted in an increase in PS I fluorescence and a decrease in the PS II fluorescence, indicating that a larger fraction of energy becomes redistributed to PS I. However, the extent of this redistribution varied: it was only small at 5°C to 20°C, but it largely increased at temperatures exceeding 20°C. This variation in the extent was related to a change in the mechanism of the state transition: at 15°C only the ‘initial’ distribution of energy was affected, while at 35°C an additional increase in the spill-over constant, k_{T (II → I)}, was included. It is assumed that under physiological conditions k_{T (II → I)} is under the control of temperature rather than of light quality, whereby in leaves adapted to high physiological temperatures, the probability of energy spill-over from closed PS II centres to PS I is enhanced. In darkened leaves, the spill-over constant has been manipulated by preincubation at different temperatures. Then, the light-induced ‘energization’ of thylakoid membranes has been tested by measuring the light-induced electrochromic absorbance change at 515 nm (and light-induced light-scattering changes) in these leaves. The flash-induced 515 nm signal as well as the initial peak during a 1 s illumination were not affected by energy distribution. However, the amplitude of the pseudo-steady-state signal (as established during 1 s illumination) was considerably enhanced in leaves in which a larger fraction of the absorbed energy is distributed to PS I at the expense of PS II excitation. The results have been interpreted in such a way that an increase in energy spill-over from PS II to PS I favours a cyclic electron transport around PS I. It is discussed that changes in energy distribution (via spill-over) may serve to maintain a suitable balance between non-cyclic and cyclic electron transport in vivo.     

Characteristics of the leaf photosynthetic machinery in broad bean grown in zinc chloride aqueous solutions

The structural and functional characteristics of bean leaves (the content of chlorophyll, the rate of oxygen production, the slow fluorescence induction, and light-induced changes in the EPR signal I from oxidized reaction centers P700+) were investigated to obtain insight into the mechanism of influence of zinc chloride on the photosynthetic apparatus. Seedlings were grown on hydroponic medium containing ZnCl2 at concentrations from 10(-7) to 10(-3) M. At low concentrations of ZnCl2, a decrease in the content of chlorophyll per one unit of leaf mass was observed, while the rate of oxygen production per chlorophyll was increased. High concentrations of ZnCl2 in the hydroponic medium caused the slowed down the plant development and inhibited the light-induced production of oxygen. The changes in biophysical characteristics of leaves the parameter FM/FT of the slow fluorescence induction, and kinetics of redox transients of P700 induced by ZnCl2 were of similar character and correlated with the changes in photosynthetic activity. The data obtained demonstrate that structural and functional changes in the photosynthetic apparatus induced by the variations of growth conditions have adaptive character.     

Light-Harvesting Function in the Diatom Phaeodactylum tricornutum: II. Distribution of Excitation Energy between the Photosystems

The distribution of excitation energy between photosystems I and II (PSI and PSII) was investigated in the marine diatom Phaeodactylum tricornutum (Bohlin) using light-induced changes in fluorescence yield and rate of modulated O₂ evolution. The intensity dependence of the fast fluorescence rise in dark adapted cells (±DCMU) suggests that light absorbed by the major antenna complex was not delivered preferentially to PSII but is more equally distributed between the photosystems. Reversible, slow fluorescence yield changes measured in the absence of DCMU were correlated with decreased initial fluorescence and rate constants for PSII photochemistry, increased variable fluorescence, alteration of the fluorescence excitation and emission spectra, and could be effected by either 510 nm (PSII) or 704 nm (PSI) light. Slow, reversible fluorescence yield changes were also observed in the presence of DCMU, but were characterized by a loss of both initial and variable fluorescence and could not be induced by PSI light. The absence of slow changes in the yield of fluorescence and rate of modulated O₂ evolution, following addition or removal of PSI background light to modulated PSII excitation, does not support regulation of excitation energy density in PSI at the expense of PSII. The results suggest that adjustments are made at the level of excitation energy transfer to the PSII reaction center which prevent prolonged loss of photosynthetic capacity. Energy distribution is regulated by ionic distributions independently of the plastoquinone pool redox state. These differences in light-harvesting function are probably a response to the aquatic light field and may account for the success of diatoms in low and variable light environments.